Amplification oligonucleotides
Disclosed herein, inter alia, are polynucleotides, supports, kits, and methods of use thereof for amplifying, immobilizing, and sequencing polynucleotides.
1. A solid support comprising a first plurality of immobilized oligonucleotides, and a second plurality of immobilized oligonucleotides, wherein each oligonucleotide of the first plurality of immobilized oligonucleotides comprises SEQ ID NO:92, and wherein each oligonucleotide of the second plurality of immobilized oligonucleotides comprises SEQ ID NO:89.
2. The solid support of claim 1 , wherein each oligonucleotide of the first plurality of immobilized oligonucleotides is covalently attached to a polymer on the solid support via a first linker.
3. The solid support of claim 1 , wherein each oligonucleotide of the second plurality of immobilized oligonucleotides is covalently attached to a polymer on the solid support via a second linker.
4. The solid support of claim 1 , wherein the solid support is a multi-well container, flow cell, particle, chip, slide, or unpatterned solid support.
5. The solid support of claim 1 , wherein the solid support further comprises a polymer, resist, and/or hydrogel.
6. The solid support of claim 1 , wherein said solid support comprises two or more wells, wherein each well comprises the first plurality of immobilized oligonucleotides and the second plurality of immobilized oligonucleotides.
7. The solid support of claim 6 , wherein each oligonucleotide of the first plurality of immobilized oligonucleotides and each oligonucleotide of the second plurality of immobilized oligonucleotides is covalently attached to a polymer within the wells.
8. A kit comprising the solid support of claim 7 .
9. The kit of claim 8 , further comprising a polymerase and a plurality of deoxynucleotides (dNTPs).
10. A microfluidic device comprising the solid support of claim 7 .
11. The solid support of claim 6 , wherein the solid support comprises about 0.2 wells to about 4.0 wells per μm 2 .
12. The solid support of claim 6 , wherein each of the wells is separated from each other by about 0.2 μm to about 2.0 μm.
13. The solid support of claim 1 , comprising about 100 oligonucleotides per μm 2 to about 1,000,000 oligonucleotides per μm 2 of said first plurality of immobilized oligonucleotides.
14. The solid support of claim 1 , comprising about 100 oligonucleotides per μm 2 to about 1,000,000 oligonucleotides per μm 2 of said second plurality of immobilized oligonucleotides.
15. The solid support of claim 1 , wherein each oligonucleotide of the first plurality of immobilized oligonucleotides is covalently attached to a polymer on the solid support via a first linker, and wherein each oligonucleotide of the second plurality of immobilized oligonucleotides is covalently attached to said polymer on the solid support via a second linker.
16. The solid support of claim 15 , wherein the first linker, the second linker, or both the first linker and the second linker comprise 8 to 16 thymine nucleotides, and wherein each linker is at the 5′ end of the immobilized oligonucleotides.
17. The solid support of claim 15 , wherein the first linker, the second linker, or both the first linker and the second linker comprises a cleavable site.
18. The solid support of claim 17 , wherein the cleavable site comprises a deoxyuracil triphosphate (dUTP), diol linker, disulfide linker, photocleavable linker, abasic site, deoxy-8-Oxo-guanine triphosphate (d-8-oxoG), methylated nucleotide, ribonucleotide, or a sequence containing a modified or unmodified nucleotide that is specifically recognized by a cleaving agent.
19. The solid support of claim 15 , wherein the first linker, the second linker, or both the first linker and the second linker comprises 1 to 5 uracil nucleotides.
20. A solid support comprising:
a first immobilized oligonucleotide comprising SEQ ID NO:92, and
a second immobilized oligonucleotide comprising SEQ ID NO:89.
21. The solid support of claim 20 , wherein the first immobilized oligonucleotide further comprises a first index sequence, and the second immobilized oligonucleotide further comprises a second index sequence.
22. The solid support of claim 20 , wherein the first immobilized oligonucleotide is covalently attached to a polymer on the solid support via a first linker, and the second immobilized oligonucleotide is covalently attached to said polymer on the solid support via a second linker.